Evaluation of Computer-Assisted Comet Assay – Repeatability and Reproducibility of Data

Abstract

Evaluation of the alkaline/neutral comet assay combined with Ag staining and computer analysis were performed and afford good repeatability and reproducibility. The present study showed that if an appropriate number of comet results (N=100) is evaluated using LUCIA image analysis, there are no statistically significant differences among operators of various levels of experience. Using public domain software CASP, the results showed a consistent tendency to provide lower average values for the studied variable (% tail DNA) compared with LUCIA software. Yet, this tendency was shown to be statistically insignificant for a majority of comets. If limited to a subsample of less damaged comets, the software comparison was statistically significant. This together with the fact that CASP was unable to process heavily damaged comets (> 90 % tail DNA) may bring certain limitations to the use of CASP combined with presented protocol. In many respects, the results of analysis using CASP are comparable with the values obtained using the LUCIA software.